CN102028646B - Thyme perfume - Google Patents
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- CN102028646B CN102028646B CN201010556741XA CN201010556741A CN102028646B CN 102028646 B CN102028646 B CN 102028646B CN 201010556741X A CN201010556741X A CN 201010556741XA CN 201010556741 A CN201010556741 A CN 201010556741A CN 102028646 B CN102028646 B CN 102028646B
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Abstract
The invention discloses thyme perfume. The thyme perfume comprises the following raw materials in part by volume: 1.92 parts of base essential oil, 0.03 to 0.10 part of thyme aromatic oil, 0 or 2 parts of essential oil emulsifier and 100 parts of anhydrous alcohol or distilled water, wherein the thyme perfume is prepared by taking the distilled water as a solvent; and 2 parts of essential oil emulsifier is added to dissolve the base essential oil in the distilled water. In the thyme perfume, the thyme aromatic oil which is extracted from thyme is used as a main component; the used thyme resource serving as a raw material is cheap and readily available, and the thyme aromatic oil serving as the main component has obvious oxidation resistance, antibacterial property and non-toxicity; the prepared perfume has good integral feeling, simple and elegant color, and harmonious and natural smell; the smell can be maintained for about 48 hours; and the thyme perfume has extremely high application value and broad market prospect.
Description
Technical field
The invention belongs to the cosmetics technical field, being specifically related to Herba thymi vulgaris aromatic oil is the perfume of main component.
Background technology
Herba thymi vulgaris is the general designation of Labiatae (Labiatae) Thymus (Thymus) plant, is one of world-renowned fragrant plant, originates in Mediterranean bank and Asia Minor one band.The Herba thymi vulgaris plant type is peculiar, short, and the flower type is little, and pattern is gorgeous, and Herb is distributed aromatic odor; The nice and cool weather of happiness, happiness light, anti-slightly shade, cold-resistant, drought-enduring, avoid wet, barren-resistant.The Herba thymi vulgaris floristics of China is not a lot, but it is wider to distribute, in the northwest, all there is distribution the North, Northeast China various places.Herba thymi vulgaris not only has higher edible, medicinal and Application in Chemical Engineering and is worth, and also has effects such as sweetening treatment, greening and environmental protection simultaneously, will be a kind of fragrant plant with very great development potentiality therefore.
The Herba thymi vulgaris aromatic-oil content is high, of fine qualities, has become the staple product in the American-European Fine Chemical Industry, and its trade is extend over the entire globe.Present stage, the aromatic oil method for distilling of Thymus plant mainly contains distillation (SD) and extraction (SDE).The analytical method of Herba thymi vulgaris aromatic oil component has a variety of, and wherein using maximum is the GC-MS analytic process.The chemical constituent of Thymus plant mainly comprises: volatile oil and nonvolatile matter, and the Herba thymi vulgaris aromatic oil composition of having reported at present reaches 47 kinds more than, and wherein main component is some alcohol, phenol, alkene class material.And nonvolatile matter mainly contains flavonoid, organic acid and many nutrients.Be rich in antioxidant contents such as thymol, carvacrol, flavone and glycoside in the Thymi Serpylli Herba extract; It is made an addition in food, beverage, health product and the cosmetics, can play anticorrosion, anti-become sour, anti aging effect and improve effects such as skin matter, hair quality.
Summary of the invention
It is main component with Herba thymi vulgaris aromatic oil that technical problem to be solved by this invention is to provide a kind of, fragrance harmony and natural and the Herba thymi vulgaris perfume of holding time long.
Solving the problems of the technologies described above the technical scheme that is adopted is that it is to be processed by following parts by volume proportion raw material:
1.92 parts of basis quintessence oils
0.03～0.10 part in Herba thymi vulgaris aromatic oil
Quintessence oil emulsifying agent 0 or 2 parts
Dehydrated alcohol or distilled water add to 100 parts
Above-mentioned basic quintessence oil is mixed and made into by Oleum menthae 0.1mL, oleum bergamottae 0.1mL, Flos Matricariae chamomillae quintessence oil 0.2mL, grapefruit quintessence oil 0.05mL, Flos Rosae Rugosae quintessence oil 0.6mL, jasmin quintessence oil 0.2mL, orange blossom quintessence oil 0.3mL, tea tree ethereal oil 0.1mL, Herba Lysimachiae foenum-graeci quintessence oil 0.1mL, cinnamon essential oil 0.1mL, rosemary ethereal oil 0.1mL, Lignum Santali Albi quintessence oil 0.02mL, Cananga odorata quintessence oil 0.05mL.
Herba thymi vulgaris perfume of the present invention is processed by following parts by volume proportion raw material:
1.92 parts of basis quintessence oils
0.07 part in Herba thymi vulgaris aromatic oil
Dehydrated alcohol adds to 100 parts
Herba thymi vulgaris perfume of the present invention also can be processed by following parts by volume proportion raw material:
1.92 parts of basis quintessence oils
0.05 part in Herba thymi vulgaris aromatic oil
2 parts of quintessence oil emulsifying agents
Distilled water adds to 100 parts
The method for preparing of above-mentioned Herba thymi vulgaris perfume is following:
1, gathers raw material
Gather Herba thymi vulgaris axis in 7～August, leaf, flower ,-18～5 ℃ of preservations or indoor are dried in the shade naturally.
2, extract Herba thymi vulgaris aromatic oil
Stem, leaf, the flower of Herba thymi vulgaris plant are put into boiler, add the distilled water of 5 times of its quality, airtight, be heated to boiling 60 minutes, condensing reflux, the buoyant aromatic oil of separating and condensing backflow upper surface obtains Herba thymi vulgaris aromatic oil.
3, preparation Herba thymi vulgaris perfume
With above-mentioned parts by volume proportion raw material mix homogeneously, in the dark-brown glass bottle of packing into, sealing, black cloth parcel was placed 15 days for 25 ℃, placed 60 days for 4 ℃, filtered, and filtrating is packed in the sealed glass jars, 4 ℃ of preservations of black cloth parcel.
The present invention is a main component with the Herba thymi vulgaris aromatic oil that from Herba thymi vulgaris, extracts, and is mixed with Herba thymi vulgaris perfume in conjunction with basic quintessence oil and dehydrated alcohol or distilled water, and the perfume integral body that obtains is felt all right, the fragrance harmony and natural, and fragrance can be kept about 48 hours.
Description of drawings
Fig. 1 is the gas-matter chromatogram of the Herba thymi vulgaris aromatic oil composition of embodiment 1 extraction.
Fig. 2 is that Herba thymi vulgaris aromatic oil is to the measured by esr technique curve chart.
Fig. 3 is the inhibitory action curve chart of Herba thymi vulgaris aromatic oil to lipid peroxidation.
The specific embodiment
To further explain of the present invention, but the invention is not restricted to these embodiment below in conjunction with accompanying drawing and embodiment.
100mL is an example with preparation Herba thymi vulgaris perfume, and is raw materials used and volume proportion is following:
Basis quintessence oil 1.92mL
Herba thymi vulgaris aromatic oil 0.07mL
Dehydrated alcohol adds to 100mL
The method for preparing of above-mentioned Herba thymi vulgaris perfume is:
1, gathers raw material
Gather Herba thymi vulgaris axis in 7～August, leaf, flower ,-18～5 ℃ of preservations or indoor are dried in the shade naturally.
2, extract Herba thymi vulgaris aromatic oil
Herba thymi vulgaris axis, leaf, flower are put into boiler, add the distilled water of 5 times of its quality, airtight, be heated to boiling 60 minutes, condensing reflux, the buoyant aromatic oil of separating and condensing backflow upper surface obtains Herba thymi vulgaris aromatic oil.
3, preparation Herba thymi vulgaris perfume
The raw materials mix of above-mentioned volume proportion is even, in the dark-brown glass bottle of packing into, sealing, black cloth parcel was placed 15 days for 25 ℃, placed 60 days for 4 ℃, filtered, and filtrating is packed in the sealed glass jars, 4 ℃ of preservations of black cloth parcel.
100mL is an example with preparation Herba thymi vulgaris perfume, and is raw materials used and volume proportion is following:
Basis quintessence oil 1.92mL
Herba thymi vulgaris aromatic oil 0.05mL
Dehydrated alcohol adds to 100mL
Its preparation method is identical with embodiment 1.
100mL is an example with preparation Herba thymi vulgaris perfume, and is raw materials used and volume proportion is following:
Basis quintessence oil 1.92mL
Herba thymi vulgaris aromatic oil 0.10mL
Dehydrated alcohol adds to 100mL
Its preparation method is identical with embodiment 1.
100mL is an example with preparation Herba thymi vulgaris perfume, and is raw materials used and volume proportion is following:
Basis quintessence oil 1.92mL
Herba thymi vulgaris aromatic oil 0.05mL
Quintessence oil emulsifying agent 2mL
Distilled water adds to 100mL
Its preparation method is identical with embodiment 1.
100mL is an example with preparation Herba thymi vulgaris perfume, and is raw materials used and volume proportion is following:
Basis quintessence oil 1.92mL
Herba thymi vulgaris aromatic oil 0.03mL
Quintessence oil emulsifying agent 2mL
Distilled water adds to 100mL
Its preparation method is identical with embodiment 1.
100mL is an example with preparation Herba thymi vulgaris perfume, and is raw materials used and volume proportion is following:
Basis quintessence oil 1.92mL
Herba thymi vulgaris aromatic oil 0.08mL
Quintessence oil emulsifying agent 2mL
Distilled water adds to 100mL
Its preparation method is identical with embodiment 1.
In order to confirm optimum process condition of the present invention and proportioning raw materials, the inventor has carried out a large amount of laboratory research tests, and various test situation are following:
Test material: Herba thymi vulgaris axis, leaf, flower are adopted in new town and country, Zhenyuan County, Gansu Province; Respectively at gathering on May 15, June 15, July 15, August 15, JIUYUE 15, October 15, adopt freezing bright Tibetan (18 ℃), the bright Tibetan of low temperature (5 ℃), the indoor three kinds of methods of drying in the shade naturally to store.Test animal is the Kunming mouse that Xi'an Communications University's medical board animal center is cultivated, and is born body weight 22～26g 45 days.
Main agents: ascorbic acid is provided by new fine chemistry industry development centre, sky, Tianjin; Potato dextrose agar solid medium (PDA solid medium) is provided by the rich Olympic star's biotechnology Co., Ltd in Beijing; Soybean lecithin is provided by Kang Puda (Wei Shi) bio tech ltd; Sulfo-barbital (TBA) is provided by SCRC Chemical Reagent Co., Ltd., Sinopharm Group; Trichloroacetic acid (TCA) is provided by the huge chemical reagent of Tianjin Dongli District factory; Vitamin E is provided by Canadian nutrition room biological medicine company limited; Hexichol is provided by Qinghai molecular biosciences reagent company for bitterness diazanyl free radical (DPPH); Supply examination strain beerwort cereuisiae fermentum, staphylococcus aureus, Aspergillus niger to provide by Shaanxi Normal University's food engineering and Microbiological Lab of nutrition science institute.
Key instrument and equipment: HP6890GC/5973MSD gas-matter coupling analyser is provided by hewlette-packard; S.SW-CJ-IF type clean work station is provided by the Shanghai medical apparatus and instruments factory of making a leapleap forward; The vertical automatic electric heating pressure steam sterilizer of LDZX-40BI type is provided by Shenan Medical Appliances Factory, Shanghai; BY-TR type optical microscope is provided by Chongqing special optical instrument company limited difficult to understand; DGX-9073B-1 type electric drying oven with forced convection is provided by Shanghai Fuma Experiment Equipment Co., Ltd.; WKY type 10-50l micropipettor is provided by Shanghai Rong Tai biochemical engineering company limited; GSP-9080MBE type water isolation type constant incubator is provided by the rich industry company limited of proving to be true after interrogation in Shanghai; MJX-250B-Z type mold incubator is provided by the rich industry company limited of proving to be true after interrogation in Shanghai; WFJ2000 type visible spectrophotometer is provided by Shanghai You Nike company limited; PL203 type electronic balance is provided by Shanghai prunus mume (sieb.) sieb.et zucc. Teller-Tuo benefit Instr Ltd..
1, the extraction of Herba thymi vulgaris aromatic oil
-18 ℃, 5 ℃ of the different times collection, the thyme and the new thyme (contrast) of gathering of the indoor three kinds of method preservations of drying in the shade are naturally respectively got 1000g, add in the boiler adding 5000g distilled water respectively; Airtight; Be heated to boiling 60 minutes, condensing reflux, the buoyant aromatic oil of separating and condensing backflow upper surface; Obtain Herba thymi vulgaris aromatic oil, calculate the aromatic oil yield by following formula (1):
Experimental result is seen table 1.
Table 1 acquisition time and method for preserving are to the influence of aromatic oil yield
| May 15 | June 15 | July 15 | August 15 | JIUYUE 15 | October 15 | |
| -18℃ | 0.11％ | 0.14％ | 0.21％ | 0.19％ | 0.15％ | 0.08％ |
| 5℃ | - | - | 0.10％ | - | - | - |
| Indoorly dry in the shade naturally | - | - | 0.15％ | - | - | - |
| Contrast | - | - | 0.22％ | - | - | - |
Annotate: contrast the fresh thyme of gathering into the same day, other are 30 days thyme of storage.
Visible by table 1, with 7, gather Herba thymi vulgaris and be advisable August, the nursery stock children is tender before July, and nursery stock is aging after JIUYUE, and the aromatic oil yield is all lower, should not gather.The Herba thymi vulgaris aromatic oil yield that adopts different method for preserving to preserve simultaneously is all higher, and wherein-18 the yield of ℃ freezing preservation Herba thymi vulgaris aromatic oil is the highest.The present invention selects-18～5 ℃ of preservations or indoor to dry in the shade naturally.
2, Herba thymi vulgaris aromatic oil composition is identified
The inventor adopts gas-matter coupling analyser that the composition of the Herba thymi vulgaris aromatic oil of the present invention's extraction is measured.GC conditions: select the HP-5MS chromatographic column for use, the service property (quality) mark is 5% benzyl polysiloxanes fused-silica capillary column (30m * 0.25mm * 0.25 μ m), and injector temperature is 200 ℃; The column compartment temperature is 60 ℃ (keeping 5min); Adopt 8 ℃/min heating rate to rise to 150 ℃ (keeping 10min), carrier gas is high-purity helium, presses before the post to be 71kPa; Flow velocity is 1mL/min, and split ratio is 15: 1.Mass spectrum condition: 280 ℃ of interface temperature, ionization mode EI (electron impact ionization), electron energy 70eV; Ion source temperature is 250 ℃, 130 ℃ of quadrupole rod temperature, and tuning manner is that standard is tuning; Mass scanning mode SCAN, solvent delay 1.5min, sweep limits is 10～550amu; Electron multiplier voltage is 1635V, and G1033A selects D.01.00NlST98 standard mass spectrum search library for use.Test result is seen Fig. 1 and table 2.
Table 2 Herba thymi vulgaris aromatic oil main chemical compositions and content
| Peak number | The chemical compound title | Molecular formula | Mass percent (%) |
| 1 | The 2-thujene | C 10H 16 | 0.43 |
| 2 | Sobrerone | C 10H 16 | 0.35 |
| 3 | Camphene | C 10H 16 | 0.47 |
| 4 | β-phellandrene | C 10H 16 | 0.34 |
| 5 | Nopinene | C 10H 16 | 0.71 |
| 6 | 3-methyl 1-isopropyl-dicyclo [3,1,0]-4-normal hexane | C 10H 16 | 0.46 |
| 7 | 4 [10]-thujenes | C 10H 16 | 0.48 |
| 8 | 1-methyl-3-cumene | C 10H 14 | 0.36 |
| 9 | Beta-myrcene | C 10H 16 | 0.34 |
| 10 | Eucalyptole | C 10H 18O | 0.78 |
| 11 | The p-P-cymene | C 10H 14 | 0.11 |
| 12 | Suitable-the 1-methyl-the 4-isopropyl-the 2-cyclohexene-1-alcohol | C 10H 18O | 0.57 |
| 13 | Instead-1-methyl-4-isopropyl-2-cyclohexene-1-alcohol | C 10H 18O | 0.99 |
| 14 | Borneolum Syntheticum | C 10H 18O | 9.04 |
| 15 | Thymol | C 12H 20O 2 | 21.60 |
| 16 | Isobornyl acetate | C 12H 20O 2 | 0.32 |
| 17 | 1-methoxyl group-3-methyl-2-cumene | C 10H 16O | 3.56 |
| 18 | Thymol | C 10H 14O | 5.03 |
| 19 | Isothymol | C 10H 14O | 4.15 |
| 20 | Acetic acid thymol ester | C 12H 16O 2 | 4.37 |
| 21 | 2-methylene-4,8,8-trimethyl-4-vinyl dicyclo [5,2,0] nonane | C 15H 24 | 6.06 |
| 22 | Germacrene-D | C 15H 24 | 4.21 |
| 23 | O-methoxy-α, α-Er Jiajibianji alcohol | C 10H 14O 2 | 0.13 |
| 24 | 2-methoxyl group-5-methyl-isopropylbenzene | C 11H 16O | 0.14 |
| 25 | 2,3,5,6-tetramethyl-1,4-benzoquinone | C 10H 12O 2 | 0.13 |
| 26 | 2-methoxyl group 4-methyl isophthalic acid-[1-Methylethyl]-benzene | C 11H 16O | 0.18 |
| 27 | Citral | C 10H 16O | 0.17 |
| 28 | Geraniol | C 10H 18O | 0.15 |
| 29 | Citral | C 10H 16O | 0.28 |
| 30 | Positive certain herbaceous plants with big flowers alcohol | C 10H 22O | 0.50 |
| 31 | Borneol acetate | C 12H 20O 2 | 0.51 |
| 32 | Instead-caryophyllene | C 15H 24 | 1.88 |
| 33 | Caryophyllene oxide | C 15H 24O | 6.89 |
| 34 | α-caryophyllene | C 15H 24 | 0.33 |
| 35 | 1,3-dimethyl cyclopentanol | C 7H 14O | 0.05 |
| 36 | Camphora | C 10H 16O | 0.07 |
| 37 | α-cadinene | C 15H 24 | 2.06 |
| 38 | γ-elemene | C 15H 24 | 1.49 |
| 39 | Carvone | C 10H 14O | 13.67 |
| 40 | Carvacrol | C 10H 14O | 24.42 |
| 41 | 1-hexyloxy-3-methyl normal hexane | C 13H 28O | 0.18 |
| 42 | 4-methyl-3-pentenals | C 6H 10O | 0.18 |
| 43 | Instead-the longleaf pine carveol | C 15H 24O | 0.2 |
| 44 | Palmic acid | C 16H 32O 2 | 0.13 |
| 45 | Humulene epoxide | C 15H 24O | 0.14 |
| 46 | δ-cadinene | C 15H 24O | 0.16 |
| 47 | Undetermined | C 15H 34O 2 | 0.13 |
| 48 | Hexahydrofarnesyl acetone | C 18H 36O | 0.11 |
| 49 | Geranyl acetone | C 13H 22O | 3.67 |
| 50 | Alloaromadendrene | C 15H 24 | 0.08 |
| 51 | Instead-the longleaf pine carveol | C 15H 24O | 0.08 |
| 52 | Undetermined | C 15H 34O 20 | 0.09 |
| Amount to | 98.51 |
Visible by Fig. 1 and table 2; 52 kinds of compositions account for 98.51% of volatile oil total amount altogether in the Herba thymi vulgaris volatile oil, and visible from qualification result, its main component is carvacrol, thymol, carvone, Borneolum Syntheticum, thymol, isothymol, 2-methylene-4; 8; 8-trimethyl-4-vinyl bicyclic nonane, acetic acid thymol ester can make an addition to them in cosmetics, anti aging effect with improve skin matter.
3, Herba thymi vulgaris aromatic oil biocidal property test
(1) preparation yeast enrichment medium
Take by weighing glucose 50g, carbamide 1g, (NH respectively
4)
2SO
47H
2O 1.0g, rose-bengal 0.03g, KH
2PO
42.5g, FeSO
47H
2O 0.1g, MgSO
47H
2O 1.0g, yeast extract 0.5g, Na
2HPO
40.5g, adding in the distilled water successively and dissolve, adding distil water adds agar 15g again to 1000mL, and heating and melting is mended distilled water to 1000mL; Use amount of substance concentration to regulate pH value to 4.5 as the NaOH of 1mol/L, filter, divide in the 200mL wide mouthed bottle of packing into as the HCl of 1mol/L and amount of substance concentration; Use the stopper jam-pack, wrapping places 100Pa sterilization 20min in the high-pressure sterilizing pot; Take out, pour in the culture dish cooled and solidified into; Be prepared into the yeast enrichment medium, be used to cultivate the beerwort cereuisiae fermentum.
(2) preparation beef extract-peptone solid medium
Take by weighing Carnis Bovis seu Bubali cream 3.0g, peptone 10g, NaCl 5.0g, agar 18g, place beaker, add the 300mL distilled water, heating makes the solid dissolving, and adding distil water is to 1000mL; Use amount of substance concentration to regulate pH value to 7.2～7.4 as the NaOH of 1mol/L, filter, divide in the 200mL wide mouthed bottle of packing into as the HCl of 1mol/L and amount of substance concentration; Use the stopper jam-pack; Wrapping places 121 ℃ of sterilization 15min in the high-pressure sterilizing pot, is cooled to 50 ℃; The examination nose end is shelved on the utensil of glass rod or other proper heights; The chamfer length of shelving is advisable with half that is no more than the test tube height overall, and cooled and solidified is prepared into the beef extract-peptone solid medium and is used to cultivate staphylococcus aureus.
(3) preparation Cha Shi culture medium
Take by weighing glucose 30g, NaNO
32.0g, K
2HPO
43H
2O 1.0g, FeSO
47H
2O 0.1g, MgSO
47H
2O0.5g, KCl 0.5g, agar 18g place beaker, add the 300mL distilled water, and heating makes the solid dissolving, and adding distil water is to 1000mL; Use amount of substance concentration to regulate pH value to 7.2 as the NaOH of 1mol/L, filter, divide in the 200mL wide mouthed bottle of packing into as the HCl of 1mol/L and amount of substance concentration; Use the stopper jam-pack, wrapping places 121 ℃ of sterilization 15min in the high-pressure sterilizing pot; Be cooled to 50 ℃, the examination nose end is shelved on the utensil of glass rod or other proper heights, the chamfer length of shelving is advisable with half that is no more than the test tube height overall; Cooled and solidified is prepared into the Cha Shi culture medium, is used to cultivate Aspergillus niger.
(4) preparation physiological saline solution
0.85g NaCl and 100mL distilled water are added in the triangular flask, tampon beyond the Great Wall, stopper outsourcing one deck kraft paper places 121 ℃ of sterilization 20min in the high-pressure sterilizing pot, and cooling is for use.
(5) preparation PDA solid medium
Take by weighing glucose 20g, agar 20g, remove the peel and smash Rhizoma Solani tuber osi 200g to pieces, place triangular flask, adding distil water stirs; Be settled to 1000mL with distilled water, use amount of substance concentration to regulate pH value to 7.6 as the NaOH of 1mol/L, place 121 ℃ of sterilization 15min in the high-pressure sterilizing pot as the HCl of 1mol/L and amount of substance concentration; Be cooled to 50 ℃, pour in the culture dish cooled and solidified into; Be prepared into the PDA solid medium, be used for the test of beerwort cereuisiae fermentum and Aspergillus niger inhibition zone.
(6) preparation supplies examination strain bacteria suspension
Get activatory staphylococcus aureus, Aspergillus niger and beerwort cereuisiae fermentum and supply the examination strain for three kinds, place the test tube that the 9mL sterile distilled water is housed respectively, fully mix, process bacteria suspension; Get the 1.0mL bacteria suspension in the test tube that the 9.0mL sterile distilled water is housed with aseptic straw, dilute successively, being mixed with spore concentration respectively is 10
6～10
7The confession examination strain bacteria suspension of individual/mL, wherein the staphylococcus aureus counting adopts colony counting method, and Aspergillus niger and beerwort cereuisiae fermentum adopt the microscope direct counting method to measure.
(7) measure inhibition zone
Be to put into Herba thymi vulgaris aromatic oil behind the filter paper dry heat sterilization of 9mm to soak 2h with diameter; Getting various each 0.15mL of confession examination strain bacteria suspension evenly coats on the corresponding solid medium; Filter paper after will in Herba thymi vulgaris aromatic oil, soaking with aseptic nipper is attached to coating and supplies on the solid medium of examination strain bacteria suspension; Each culture dish pastes 3; Aseptic filter paper behind the dry heat sterilization learn from else's experience simultaneously as blank, the beef extract-peptone solid medium of coating staphylococcus aureus bacteria suspension is cultivated 24h for 37 ℃, 28 ℃ of cultivations of PDA solid medium 48h of coating Aspergillus niger bacteria suspension and beerwort cereuisiae fermentum bacteria suspension.Measure the antibacterial circle diameter of filter paper, the result gets the meansigma methods of three repeated trials, and test result is seen table 3.Estimate fungistatic effect according to inhibition zone, the inhibition zone criterion of antibiotics is: for responsive type, being the medium sensitivity type during 10～15mm during antibacterial circle diameter＞15mm, is low responsive type during 7～9mm, and no fungistatic effect person is a non-sensitive type.
Table 3 Herba thymi vulgaris aromatic oil is to supplying the bacteriostasis of examination strain
| Supply the examination strain | Antibacterial circle diameter (mm) |
| Aspergillus niger | 21.4 |
| Staphylococcus aureus | 14.9 |
| The beerwort cereuisiae fermentum | 18.4 |
| Blank | 9.48 |
Visible by table 3; The antibacterial circle diameter of Aspergillus niger, beerwort cereuisiae fermentum is all more than 15mm; Belong to responsive type, staphylococcus aureus belongs to the medium sensitivity type, explains that Herba thymi vulgaris aromatic oil all has stronger inhibitory action to staphylococcus aureus, Aspergillus niger and the three kinds of strains of beerwort cereuisiae fermentum that supply examination; And the inhibitory action to Aspergillus niger is best, secondly is beerwort cereuisiae fermentum and staphylococcus aureus.
(8) pH value is to Herba thymi vulgaris aromatic oil bacteriostatic test
The pH value that uses amount of substance concentration to regulate the beef extract-peptone solid medium as NaOH solution and the amount of substance concentration of 0.5mol/L as the HCl solution of 0.5mol/L is 5,6,7,8; Supplying examination strain bacteria suspension is the staphylococcus aureus bacteria suspension; Adopt the said determination method to measure the size of inhibition zone; More different pH value are to the influence of Herba thymi vulgaris aromatic oil fungistatic effect, and the result sees table 4.
The different pH value of table 4 are to the influence of Herba thymi vulgaris aromatic oil bacteriostasis
| |
5 | 6 | 7 | 8 |
| Antibacterial circle diameter/mm | 18.2 | 17.4 | 16.08 | 15.3 |
Visible by table 4, different pH value have stronger influence to the biocidal property of Herba thymi vulgaris aromatic oil, and the bacteriostasis of low pH is stronger, and pH value is 5,6 o'clock, and thalline has stopped growing fully; PH value is 7,8 o'clock, and with the increase of pH value, inhibition zone has the trend that reduces.This fungistatic effect that shows Herba thymi vulgaris aromatic oil is comparatively strong under acid condition, and the increase with pH value begins to weaken to some extent under neutral and alkali condition.Hence one can see that, reduces the bacteriostasis that pH can strengthen Herba thymi vulgaris aromatic oil.
(9) the antibacterial heat stabilization test of Herba thymi vulgaris aromatic oil
With Herba thymi vulgaris aromatic oil respectively at 80 ℃, 100 ℃, 121 ℃ following heat treatment 15min; Be chilled to room temperature; Supplying examination strain bacteria suspension is that pH value is 7.0 staphylococcus aureus strain bacteria suspension, adopts the method for testing of above-mentioned inhibition zone to measure its fungistatic effect respectively, and the result sees table 5.
The influence of table 5 Herba thymi vulgaris aromatic oil heat treated biocidal property
| Temperature/ |
37℃ | 80℃ | 90℃ | 121℃ |
| Antibacterial circle diameter/mm | 15.9 | 11.5 | 11.8 | 11.3 |
Visible by table 5, Herba thymi vulgaris aromatic oil after heat treatment has certain abated effect to the fungistatic effect of staphylococcus aureus, and when heating temperature reached more than 80 ℃, fungistatic effect changed not obvious.
4, Herba thymi vulgaris aromatic oil oxidation resistance test
(1) hexichol is for bitterness diazanyl free radical method (DPPH method)
1. prepare sample solution: get 10 test tubes, adding mass fraction successively according to the order of table 6 is that 95% alcoholic solution, amount of substance concentration are 2 * 10
-41, the 1 diphenyl picryl phenylhydrazine aqueous solution (DPPH aqueous solution) of mol/L, the Herba thymi vulgaris aromatic oil solution of different volumes (it is formulated that 4.0mL Herba thymi vulgaris aromatic oil sample uses mass fraction to be that 95% alcoholic solution is settled to 100mL).
Table 6 is measured Herba thymi vulgaris aromatic oil non-oxidizability reagent preparation table with the DPPH method
2. prepare standard specimen: with 1.0mL amount of substance concentration is 2 * 10
-4The DPPH solution of mol/L and 2.0mL mass fraction are 95% alcoholic solution mixing.
3. prepare control sample: the Herba thymi vulgaris aromatic oil solution in the sample solution is used isopyknic vitamin E solution (it is formulated to use mass fraction to be that 95% alcoholic solution is settled to 100mL the 4.0g vitamin E) replacement respectively, and other raw materials and addition thereof are identical with sample solution.
Sample solution and corresponding control sample, standard specimen are placed 30min respectively, place visible spectrophotometer, measure absorbance at the 517nm place, calculate its suppression ratio (promptly removing the ability of free radical) by following formula (2):
I=[(A
Standard-A
Sample)/A
Standard] * 100% (2)
I representes suppression ratio in the formula, A
StandardExpression standard specimen absorbance, A
SampleThe absorbance of expression sample solution or control sample.Experimental result is seen Fig. 2.
Visible by Fig. 2, the ability that Herba thymi vulgaris aromatic oil is removed free radical is different under different concentration, but whole linear.The ability of removing free radical as the vitamin E of controlled trial increases along with the increase of concentration, and is also linear.The ability of Herba thymi vulgaris aromatic oil removing free radical is more intense, and between 0.6～0.9, when the interpolation sample size was 0.6mL (test tube numbering 1), Herba thymi vulgaris aromatic oil was removed the free radical ability and obviously is better than vitamin E especially.
(2) lipid peroxidation method
1. prepare sample solution: get 7 test tubes, according to table 7 order add the Herba thymi vulgaris aromatic oil solution (it is formulated that 4.0mL Herba thymi vulgaris aromatic oil is settled to 100mL with sugar esters) of 0.8mL soybean lecithin buffer solution (it is that 7.4 amount of substance concentration are that the PBS of 0.1mol/L is formulated that the 0.03g soybean lecithin is dissolved in 100mL pH), different volumes successively, the pH of different volumes is that 7.4 amount of substance concentration are PBS (PBS buffer solution) and the 0.2mL FeSO of 0.1mol/L
4-Vc solution (0.141g FeSO
4Be dissolved in the 10mL distilled water formulated with 0.152gVc).
Table 7 is measured Herba thymi vulgaris aromatic oil non-oxidizability reagent preparation table with the lipid peroxidation method
2. prepare standard specimen: with soybean lecithin buffer solution, the 0.2mL FeSO of 0.8mL
4-Vc solution, 2.0mL PBS buffer solution mixes.
3. prepare control sample solution: the Herba thymi vulgaris aromatic oil solution in the sample solution is used isopyknic vitamin E solution (it is formulated that the 4.0g vitamin E is settled to 100mL with sugar esters) replacement respectively; Other raw materials and addition thereof are identical with sample solution, are mixed with control sample solution.
Sample solution, control sample, standard specimen are heated 15min in 98 ℃ of water-baths, take out, cooling, the supernatant is got in centrifugalize, places visible spectrophotometer, measures absorbance at the 532nm place, calculates its suppression ratio according to formula (2).Experimental result is seen Fig. 3.
Visible by Fig. 3, Herba thymi vulgaris aromatic oil has stronger lipoid peroxidization resistant.When the addition of Herba thymi vulgaris aromatic oil solution was 0.4～1.6mL, its oxidation resistance changed little, and its lipid peroxidation suppression ratio is 0.4%～0.6%.When Herba thymi vulgaris aromatic oil solution addition was 0.4mL (being test tube 7), its suppression ratio explained that apparently higher than vitamin E the Herba thymi vulgaris aromatic oil inhibition lipid peroxidation ability of low concentration is better than vitamin E.
5, Herba thymi vulgaris aromatic oil emergency toxicology test
(1) white mice 0%～100% deadly consumption trial test
The dosage range of white mice fatality rate 0%～100% just is decided to be 1; 0mL/kg～5.0mL/kg; The experiment white mice is divided into 5 groups by various dose; Every group 4 (male and female half and half), disposable filling stomach, every group of filling stomach dosage is: 1.00mL/kg, 2.00mL/kg, 3.00mL/kg, 4.00mL/kg, 5.00mL/kg.
Experiment is found white mice after 5.00mL/kg Herba thymi vulgaris aromatic oil is irritated stomach, occur palpitating speed, heart rate irregular, send undesired cackle, balance variation, wave on foot, paces are unstable, skin presents unusual bluish violet (the afterbody blood vessel is particularly evident), ear hyperemia, eyeball congestion, tic of the limbs, moving difficulty, until the symptom of respiratory arrest.
(2) Herba thymi vulgaris aromatic oil is to the white mice acute toxinology experiment
Adopt the improvement karber's method, according to the result of test (1), the dosage range of white mice fatality rate 0%～100% just is decided to be 1.50mL/kg～4.00mL/kg, the test white mice is divided into 8 groups by various dose, 10 every group (male and female half and half), disposable filling stomach.By r=lg
-1[(lgb-lga)/n-1], wherein r is the group distance, and b is 100% fatal dose, and a is 0% fatal dose, and n is the group number.Through calculating r ≈ 1.15.Divide into groups according to r, irritate stomach dosage for every group and just be made as: 1.50mL/kg, 1.73mL/kg, 1.98mL/kg, 2.28mL/kg, 2.62mL/kg, 3.02mL/kg, 3.47mL/kg, 4.00mL/kg.Observe 1 every day after giving mice lavage, and observed and recorded draws LD (white mice lethal dose) and LD 15 days poisoning manifestations, death toll and death times
50(white mice half lethal dose).For the white mice that do not cause death, after the off-test, take by weighing body weight, putting to death also, the tissue of major organs such as its heart of anatomic observation, liver, spleen, lung, kidney stomach function regulating changes record.The result sees table 8.
Table 8 white mice Herba thymi vulgaris aromatic oil is irritated excess of the stomach and is tested the result
| Experimental group | Irritate stomach dosage (mL/kg) | Death toll (only) | Mortality rate (%) |
| 1 | 1.50 | 0 | 0 |
| 2 | 1.73 | 0 | 0 |
| 3 | 1.98 | 2 | 20 |
| 4 | 2.28 | 2 | 20 |
| 5 | 2.62 | 3 | 30 |
| 6 | 3.02 | 5 | 50 |
| 7 | 3.47 | 8 | 80 |
| 8 | 4.00 | 10 | 100 |
Visible by table 8, the acute toxicity lethal dose (LD) of Herba thymi vulgaris aromatic oil is 4.00mL/kg, half lethal dose (LD
50) be 3.02mL/kg.
(press the half lethal dose LD of material according to American Academy of Sciences's toxicant grading standard
50Value is divided): 0 grade of avirulence, LD
50＞15.0g/kg; 1 grade of actual avirulence, 5.0g/kg＜LD
50＜15.0g/kg; 2 grades of slight toxicity, 0.5g/kg＜LD
50＜5.0g/kg; 3 grades of moderate toxicity, 50.0mg/kg＜LD
50＜500.0mg/kg; 4 grades of toxicity, LD
50＜50.0mg/kg.The toxic degree of Herba thymi vulgaris aromatic oil is 2 grades, is the slight toxic product of a kind of tool, LD
50Be the 3.02mL/kg body weight, the LD value is the 4.00mL/kg body weight.
(3) Herba thymi vulgaris aromatic oil is to the influence of white mice main organs
Use dosage as 1.50mL/kg and 1.73mL/kg to mice lavage after, slight poisoning symptom appears in white mice in the 24h, symptom fades away behind the 24h; To 15 days; Put to death all white mice that do not cause death, dissect, observe internal organs such as the white mice heart, liver, spleen, lung, kidney stomach function regulating; Warp is compared with the matched group white mice of not irritating Herba thymi vulgaris aromatic oil, and not seeing has abnormal change.
6, Herba thymi vulgaris perfume test preparation
(1) prepares basic quintessence oil
With Oleum menthae 0.1mL, oleum bergamottae 0.1mL, Flos Matricariae chamomillae quintessence oil 0.2mL, grapefruit quintessence oil 0.05mL, Flos Rosae Rugosae quintessence oil 0.6mL, jasmin quintessence oil 0.2mL, orange blossom quintessence oil 0.3mL, tea tree ethereal oil 0.1mL, Herba Lysimachiae foenum-graeci quintessence oil 0.1mL, cinnamon essential oil 0.1mL, rosemary ethereal oil 0.1mL, Lignum Santali Albi quintessence oil 0.02mL, Cananga odorata quintessence oil 0.05mL mix homogeneously, be mixed with basic quintessence oil.
(2) preparation alcohol-based Herba thymi vulgaris perfume
In basic quintessence oil, add Herba thymi vulgaris aromatic oil 0.05mL, 0.06mL, 0.07mL, 0.08mL, 0.09mL, 0.10mL respectively, be settled to 100mL with dehydrated alcohol, paraffin sealing left standstill 60 days in shady and cool lucifuge place, and solvent, solute are fully mixed.
6 testers of random arrangement to the Herba thymi vulgaris perfume of different formulations preparation, feel that from the holding time of Herba thymi vulgaris aromatic oil local flavor, fragrance, color and perfume integral body several aspects estimate respectively, and total points 30 minutes is perfume marking, and standards of grading are seen table 9.Get the meansigma methods of 6 tester's scorings, the sense organ evaluating result of alcohol-based Herba thymi vulgaris perfume is seen table 10.
Table 9 Herba thymi vulgaris perfume sense organ test and appraisal project and standards of grading
Table 10 alcohol-based Herba thymi vulgaris perfume sense organ evaluating result
| Herba thymi vulgaris aromatic oil addition (mL) | The perfume local flavor | Hold time | The perfume color | The whole sensation of fragrance | Total points |
| 0.05 | 3 | 4 | 6 | 4 | 17 |
| 0.06 | 4 | 8 | 10 | 5 | 27 |
| 0.07 | 5 | 9 | 10 | 5 | 29 |
| 0.08 | 4 | 8 | 9 | 4 | 25 |
| 0.09 | 4 | 9 | 3 | 3 | 19 |
| 0.10 | 3 | 9 | 2 | 3 | 13 |
Visible by table 10, in alcohol-based Herba thymi vulgaris perfume recipe, when the addition of Herba thymi vulgaris aromatic oil is 0.07mL; The perfume PTS is the highest, and its integral body feels best, and this prescription both can obviously be experienced thymic peat-reek; Can not make the too outstanding perfume whole structure that influences of its taste again; Perfume taste associative perception feel harmony and natural, color and luster is simple and elegant, and fragrance can be kept about 48h.Along with the increase of Herba thymi vulgaris aromatic oil addition, the perfume color and luster is deepened gradually, whole local flavor from simple and elegant until intense stimulus.The present invention selects the best 0.07mL/100mL of being of the addition of Herba thymi vulgaris aromatic oil in the alcohol-based Herba thymi vulgaris perfume.
(3) the water base Herba thymi vulgaris aromatic oil of preparation
In basic quintessence oil, add Herba thymi vulgaris aromatic oil 0.03mL, 0.04mL, 0.05mL, 0.06mL, 0.07mL, 0.08mL respectively; Add quintessence oil emulsifying agent 2.0mL, be settled to 100mL with distilled water, paraffin sealing; Left standstill 60 days in shady and cool lucifuge place, solvent, solute are fully mixed.The sense organ water base Herba thymi vulgaris perfume of testing and assessing, evaluation standard is identical with table 9, and evaluating result is seen table 11.
The water base Herba thymi vulgaris perfume of table 11 sense organ evaluating result
Visible by table 11, in water base Herba thymi vulgaris perfume recipe, when Herba thymi vulgaris aromatic oil addition was 0.05mL, the total points of Herba thymi vulgaris perfume was the highest, and its integral body feels best.Because need various quintessence oils are soluble in water; Add quintessence oil emulsifying agent 2.0mL/100mL in the water base Herba thymi vulgaris perfume process for preparation; Make the whole color and luster of water base Herba thymi vulgaris perfume present the milky of variable concentrations, on the permeability of vision, slightly poorer than alcohol-based Herba thymi vulgaris perfume.Simultaneously, the dissolubility of water base Herba thymi vulgaris perfume also is worse than alcohol-based Herba thymi vulgaris perfume.Experiment draws, and adds equivalent Herba thymi vulgaris aromatic oil, and the abnormal smells from the patient of water base Herba thymi vulgaris perfume is superior to alcohol-based Herba thymi vulgaris perfume, the slightly weak point but fragrance is held time.
7, after-ripening ageing
The alcohol-based Herba thymi vulgaris perfume for preparing is packed in the amber bottle, and room temperature is placed under the dark condition, carries out the after-ripening ageing test, and the result sees table 12.
Table 12 alcohol-based Herba thymi vulgaris perfume after-ripening ageing sense organ evaluating result
| Digestion time (my god) | The essence local flavor | Hold time | The perfume color | The whole sensation of fragrance | Total points |
| 10 | 3 | 5 | 6 | 3 | 17 |
| 20 | 3 | 6 | 8 | 3 | 20 |
| 40 | 4 | 7 | 9 | 5 | 25 |
| 60 | 5 | 9 | 10 | 5 | 29 |
| 80 | 5 | 9 | 10 | 4 | 28 |
| 100 | 5 | 9 | 10 | 5 | 29 |
Visible by table 12, Herba thymi vulgaris perfume after-ripening ageing is more than 60 days, the essence local flavor, hold time, perfume color, whole structure be all good.
Claims (3)
1. Herba thymi vulgaris perfume is characterized in that it is processed by following parts by volume proportion raw material:
Above-mentioned basic quintessence oil is mixed and made into by Oleum menthae 0.1mL, oleum bergamottae 0.1mL, Flos Matricariae chamomillae quintessence oil 0.2mL, grapefruit quintessence oil 0.05mL, Flos Rosae Rugosae quintessence oil 0.6mL, jasmin quintessence oil 0.2mL, orange blossom quintessence oil 0.3mL, tea tree ethereal oil 0.1mL, Herba Lysimachiae foenum-graeci quintessence oil 0.1mL, cinnamon essential oil 0.1mL, rosemary ethereal oil 0.1mL, Lignum Santali Albi quintessence oil 0.02mL, Cananga odorata quintessence oil 0.05mL.
2. according to the described Herba thymi vulgaris perfume of claim 1, it is characterized in that it is processed by following parts by volume proportion raw material:
1.92 parts of basis quintessence oils
0.07 part in Herba thymi vulgaris aromatic oil
Dehydrated alcohol adds to 100 parts.
3. according to the described Herba thymi vulgaris perfume of claim 1, it is characterized in that it is processed by following parts by volume proportion raw material:
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